| GAL4 DNA-Binding Domain (GAL4 DBD) |
Serves as a control DBD in yeast two-hybrid assays; binds GAL4-UAS sequences to initiate transcription when paired with AD fusion. |
BBa_25MD9GE0 |
| LexA DNA-Binding Domain (LexA DBD) |
Core DNA-binding region from E. coli LexA repressor; used as bait domain in LexA-based Y2H system to avoid cross-talk with yeast GAL4 system. |
BBa_K5477045 |
| ESR1 (ERα) Full-Length CDS |
Encodes human estrogen receptor α; mediates ligand-dependent dimerization upon binding estradiol or BPA. |
BBa_K2365510 |
| Flexible Peptide Linker |
Connects functional protein domains (e.g., LexA-DBD and ESR1 LBD) without interfering with folding or activity. |
BBa_K3712011 |
| LexA Operator Sites (LexAop) |
DNA recognition sites for LexA DBD; placed upstream of reporter genes (e.g., lacZ) to control transcriptional output. |
BBa_K5154000 |
| Reporter: lacZ (β-Galactosidase) |
Reporter enzyme that produces a colorimetric signal (yellow/blue) to indicate successful ESR1 dimerization in yeast. |
BBa_25ZHGV3T |
| Reporter/Selection Marker: LEU2 |
Yeast auxotrophic marker allowing growth in leucine-deficient media; part of the reporter plasmid system. |
BBa_K5159020 |
| pGilda (LexA-BD Vector) |
Bait plasmid carrying LexA DNA-binding domain under GAL1 promoter; used for expressing LexA-ESR1 fusion proteins. |
BBa_25TNIQC1 |
| pB42AD / pJG4-5 (B42 AD Vector) |
Prey plasmid expressing B42 activation domain fusions; interacts with bait proteins to activate reporter transcription. |
BBa_25QXAFSB |
| pGEM-T (TA Cloning Vector) |
PCR product cloning vector with T-overhangs; used as ESR1 gene source before subcloning into Y2H plasmids. |
BBa_252BAXQ8 |
| EGY48 Yeast Strain |
Host yeast strain for LexA-based Y2H; contains chromosomal LexAop-lacZ reporter and auxotrophic markers. |
BBa_250R8SL4 |
| EcoRI Restriction Site |
Restriction enzyme site (GAATTC) used for cloning ESR1 fragments into both plasmids. |
BBa_255XOW4U |
| BamHI Restriction Site |
Restriction enzyme site (GGATCC) used for directional cloning in pGilda vector. |
BBa_252D4G09 |
| ESR1 Mutant FW Primer |
Forward primer for introducing site-specific mutations into ESR1 LBD (Y519, G523, N524, T529, L535). |
BBa_2555K2IP |
| ESR1 Mutant 300 (RV) Primer |
Reverse primer targeting ESR1 300+ region for amplification and mutation validation. |
BBa_25QB456Z |